Tgf-beta inhibitors and therapeutic use thereof

Abstract

The present disclosure provides TGFβ inhibitor therapy for treating immunosuppressive conditions, cancer, and fibrosis, either as a monotherapy or combination/adjunct therapy. Selection of suitable therapy and patients who are likely to benefit from such therapy are also disclosed, as well as methods of treating cancer and fibrosis and methods of predicting and monitoring therapeutic response. Related compositions, methods and therapeutic use are also disclosed.

1 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of cancer in a subject, comprising:
 (i) determining a level of circulating TGFβ in the subject prior to administering a TGFβ antibody or antigen-binding fragment thereof; 
 (ii) administering to the subject a therapeutically effective amount of the TGFβ antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1001 (H-CDR1), SEQ ID NO: 1002 (H-CDR2), and SEQ ID NO: 1003 (H-CDR3) and three light chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1004 (L-CDR1), SEQ ID NO: 1005 (L-CDR2), and SEQ ID NO: 1006 (L-CDR3), as defined by the IMTG numbering system, wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1007 and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 1008 or amino acid sequences 90% identical thereto; 
 (iii) determining a level of circulating TGFβ in the subject after administration; and 
 wherein treatment increases the level of circulating TGFβ in the subject, and wherein the circulating TGFβ level is determined or has been determined by processing a blood sample from the subject below room temperature in a sample tube comprising an anticoagulant. 
 
     
     
         2 . Use of a TGFβ antibody or antigen-binding fragment thereof in determining therapeutic efficacy in a subject being treated for cancer, comprising:
 (i) determining a level of circulating MDSC in a blood sample collected from the subject prior to administering a TGFβ antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1001 (H-CDR1), SEQ ID NO: 1002 (H-CDR2), and SEQ ID NO: 1003 (H-CDR3) and three light chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1004 (L-CDR1), SEQ ID NO: 1005 (L-CDR2), and SEQ ID NO: 1006 (L-CDR3), as defined by the IMTG numbering system, wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1007 and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 1008 or amino acid sequences 90% identical thereto; and 
 (ii) determining a level of circulating MDSC in a blood sample collected from the subject after the administration; 
 wherein a decrease in the circulating MDSC level after the administration as compared to before the administration indicates therapeutic efficacy; 
 wherein, optionally, the decrease is at least 10%. 
 
     
     
         3 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of cancer in a subject, comprising:
 (i) determining a level of circulating MDSC in a blood sample collected from the subject prior to administering a TGFβ antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1001 (H-CDR1), SEQ ID NO: 1002 (H-CDR2), and SEQ ID NO: 1003 (H-CDR3) and three light chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1004 (L-CDR1), SEQ ID NO: 1005 (L-CDR2), and SEQ ID NO: 1006 (L-CDR3), as defined by the IMTG numbering system, wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1007 and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 1008 or amino acid sequences 90% identical thereto; and 
 (ii) determining a level of circulating MDSC in a blood sample collected from the subject after the administration; 
 (iii) continuing to administer TGFβ antibody or antigen-binding fragment thereof if a decrease in the circulating MDSC level after the administration as compared to before the administration is detected; 
 wherein, optionally, the decrease is at least 10%. 
 
     
     
         4 . The use of  claim 2 or 3 , wherein determining a level of circulating MDSC comprises determining a level of circulating mMDSC and/or a level of circulating gMDSC;
 wherein the circulating mMDSC level is determined by measuring the level of cells expressing CD11b+, HLADR−/low, CD14+, CD15−, CD33+/high, and CD66b and a decrease of at least 10% in the level of circulating mMDSC after the administration as compared to before the administration indicates therapeutic efficacy; and/or   wherein the circulating gMDSC level is determined by measuring the level of cells expressing CD11b+, HLADR−, CD14−, CD15+, CD33+/low, and CD66+ and a decrease of at least 10% in the level of circulating gMDSC after the administration as compared to before the administration indicates therapeutic efficacy.   
     
     
         5 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of a disease associated with dysregulation of the extracellular matrix in a subject, comprising:
 administering to the subject a therapeutically effective amount of the TGFβ antibody or antigen-binding fragment thereof; 
 wherein the therapeutically effective amount is selected to cause a change in the expression of one or more genes selected from ACTB, pSmad2, HPRT, CCL2, COL1a1, COL3a1, CTGF, CD68, TIMP1, LCN2, HAVCR1, TNFα, FN1, IL-1b, IL-6, LOX, ITGA11, LOXL2, ACTA2, LRRC32, LTBP1, LTBP3, MMP2, MMP9, NRROS, SERPINE1, TGFb1, TGFb2, TGFb3, and THBS1, preferably wherein the one or more genes comprises ACTA2, Col1a1, COL3a1, and/or pSmad2. 
 
     
     
         6 . The use of  claim 5 , wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1001 (H-CDR1), SEQ ID NO: 1002 (H-CDR2), and SEQ ID NO: 1003 (H-CDR3) and three light chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1004 (L-CDR1), SEQ ID NO: 1005 (L-CDR2), and SEQ ID NO: 1006 (L-CDR3), as defined by the IMTG numbering system, wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1007 and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 1008 or amino acid sequences 90% identical thereto and the disease is cancer. 
     
     
         7 . The use of  claim 5 or 6 , wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of GFTFADYA (SEQ ID NO: 276); ISGSGAAT (SEQ ID NO: 282); VSSGHWDYD (SEQ ID NO: 287); and three light chain complementarity determining regions comprising amino acid sequences of QSISSY (SEQ ID NO: 279); AASGLES (SEQ ID NO: 284); and, QQTYGVPLT (SEQ ID NO: 285), wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:297, and a light chain variable region comprising an amino acid sequence of SEQ ID NO:298 or amino acid sequences 90% identical thereto and the disease is fibrosis. 
     
     
         8 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at 240-3000 mg per dose every 2 weeks or every 3 weeks. 
     
     
         9 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 1000 mg every 2 weeks or at a dose of 1500 mg every 3 weeks. 
     
     
         10 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 80, 240, 1600, 2400, or 3000 mg every 2 weeks. 
     
     
         11 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 80, 240, 1600, 2400, or 3000 mg every 3 weeks. 
     
     
         12 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 800, 1600, 2000, or 3000 mg every 3 weeks. 
     
     
         13 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 1500 mg every 3 weeks. 
     
     
         14 . The use of  any one of the preceding claims , wherein the TGFβ antibody or antigen-binding fragment thereof is administered at a dose of 1000 mg every 2 weeks. 
     
     
         15 . The use of  any one of the preceding claims , comprising administering a checkpoint inhibitor therapy concurrently (e.g., simultaneously), separately, or sequentially with the TGFβ antibody or antigen-binding fragment thereof, wherein the checkpoint inhibitor is optionally an anti-PD-1 antibody, anti-PD-L1 antibody, anti-CTLA-4-antibody, anti-LAG3 antibody, or an antigen-binding fragment thereof. 
     
     
         16 . The use of  any one of the preceding claims , wherein the cancer comprises a solid tumor. 
     
     
         17 . The use of  any one of the preceding claims , wherein the solid tumor is selected from the group consisting of melanoma (e.g., metastatic melanoma), renal cell carcinoma, triple-negative breast cancer, HER2-positive breast cancer, colorectal cancer (e.g., microsatellite stable-colorectal cancer), lung cancer (e.g., metastatic non-small cell lung cancer, small cell lung cancer), esophageal cancer, pancreatic cancer, bladder cancer, kidney cancer (e.g., transitional cell carcinoma, renal sarcoma, and renal cell carcinoma (RCC), including clear cell RCC, papillary RCC, chromophobe RCC, collecting duct RCC, or unclassified RCC), uterine cancer, prostate cancer, stomach cancer (e.g., gastric cancer), head and neck squamous cell cancer, urothelial carcinoma, hepatocellular carcinoma, and thyroid cancer. 
     
     
         18 . The use of  any one of the preceding claims , wherein the cancer has an immune excluded phenotype, e.g., characterized by containing less than 5% intratumor CD8+ cells and greater than 5% margin CD8+ cells as assessed by an immunohistochemistry analysis capable of detecting individual tumor nest(s) within the tumor. 
     
     
         19 . The use of  any one of the preceding claims , wherein the cancer is characterized by having greater than 50% tumor area comprising tumor nest(s) comprising lower levels of CD8+ cells inside the tumor nest(s) relative to levels of CD8+ cells outside of the tumor nest, e.g., less than 5% CD8+ cells inside the tumor nest(s) and greater than 5% CD8+ cells outside the tumor nest(s), e.g., in the margin. 
     
     
         20 . The use of  any one of the preceding claims , wherein circulating TGFβ levels after administration as is increased at least 1.5-fold, at least 2-fold, at least 2.5-fold, at least 3-fold, at least 4-fold, at least 5-fold, or more compared to before administration. 
     
     
         21 . The use of  any one of the preceding claims , wherein the subject does not exhibit one or more of cardiac toxicities, neurotoxicities, or ocular toxicities after administration. 
     
     
         22 . The use of  any one of the preceding claims , wherein the subject does not exhibit one of more of fatigue, abdominal pain, back pain, nausea, vomiting, hypoxia, dyspnoea, hypokalaemia, hypomagnesaemia, or rash maculo-papular after administration. 
     
     
         23 . The use of  any one of the preceding claims , wherein the subject does not exhibit one of more of appetite decrease, ALT increase, AST increase, lipase increase, troponin increase, blood creatine increase, after administration as compared to before administration. 
     
     
         24 . The use of  any one of the preceding claims , wherein a dosage of the TGFβ antibody or antigen-binding fragment achieves stable disease, e.g., stable for great than or equal to 100, 150, 200, or 250 days. 
     
     
         25 . The use of  any one of the preceding claims , wherein a dosage of the TGFβ antibody or antigen-binding fragment achieves stable disease for greater than or equal to 153 days with tumor regressions. 
     
     
         26 . The use of  any one of the preceding claims , wherein a dosage of the TGFβ antibody or antigen-binding fragment achieves stable disease for greater than or equal to 245 days with tumor regressions. 
     
     
         27 . The use of  any one of the preceding claims , wherein a dosage of the TGFβ antibody or antigen-binding fragment achieves a partial response, e.g., as measured by RECIST1.1 PR. 
     
     
         28 . The use of  any one of the preceding claims , wherein a dosage of the TGFβ antibody or antigen-binding fragment exhibits clearance at steady state between 0.0164 to 0.225 L/h and/or volume distribution at steady state between 4.21 to 6.85 L, preferably wherein the dosage comprises at least 800 mg of the TGFβ antibody or antigen-binding fragment. 
     
     
         29 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of cancer in a subject, comprising:
 (i) determining a level of circulating TGFβ in the subject prior to administering a TGFβ antibody or antigen-binding fragment thereof; 
 (ii) administering to the subject a first dose of the TGFβ antibody or antigen-binding fragment thereof, wherein the antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1001 (H-CDR1), SEQ ID NO: 1002 (H-CDR2), and SEQ ID NO: 1003 (H-CDR3) and three light chain complementarity determining regions comprising amino acid sequences of SEQ ID NO: 1004 (L-CDR1), SEQ ID NO: 1005 (L-CDR2), and SEQ ID NO: 1006 (L-CDR3), as defined by the IMTG numbering system, wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO: 1007 and a light chain variable region comprising an amino acid sequence of SEQ ID NO: 1008 or amino acid sequences 90% identical thereto; 
 (iii) determining a level of circulating TGFβ in the subject after administration; and 
 (iv) administering a second dose of the TGFβ antibody or antigen-binding fragment thereof to the subject if the level of circulating TGFβ is elevated, 
 wherein determining the level of TGFβ comprises processing a blood sample from the subject below room temperature in a sample tube comprising an anticoagulant. 
 
     
     
         30 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of fibrosis in a subject, comprising:
 administering a therapeutically effective amount of a TGFβ antibody or antigen-binding fragment thereof to the subject as a loading dose/maintenance dose regimen, wherein the TGFβ antibody or antigen-binding fragment thereof inhibits TGFβ1 but does not inhibit one or both of TGFβ2 and/or TGFβ3, 
 thereby treating fibrosis in the subject. 
 
     
     
         31 . Use of a TGFβ antibody or antigen-binding fragment thereof in the treatment of fibrosis in a subject, comprising steps of:
 administering a therapeutically effective amount of a TGFβ antibody or antigen-binding fragment thereof to the subject, wherein the TGFβ inhibitor inhibits TGFβ1 but does not inhibit one or both of TGFβ2 and/or TGFβ3, in an amount effective to
 (i) reduce the amount of collagen present in a fibrotic tissue in the subject after administration, as compared to the amount of collagen present in the fibrotic tissue in the subject prior to administration; 
 (ii) reduce the amount of new collagen synthesis in a fibrotic tissue in the subject after administration, as compared to the amount of new collagen synthesis present in the fibrotic tissue in the subject prior to administration; and/or 
 (iii) reduce the amount of phosphorylated Smad2 in a fibrotic tissue in the subject after administration, as compared to the amount of phosphorylated Smad2 present in the fibrotic tissue in the subject prior to administration; 
 
 thereby treating fibrosis in the subject. 
 
     
     
         32 . The use of  claim 30 or 31 , wherein the TGFβ antibody or antigen-binding fragment thereof comprises three heavy chain complementarity determining regions comprising amino acid sequences of GFTFADYA (SEQ ID NO: 276); ISGSGAAT (SEQ ID NO: 282); VSSGHWDYD (SEQ ID NO: 287); and three light chain complementarity determining regions comprising amino acid sequences of QSISSY (SEQ ID NO: 279); AASGLES (SEQ ID NO: 284); and, QQTYGVPLT (SEQ ID NO: 285), wherein optionally the TGFβ inhibitor comprises an isolated antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence of SEQ ID NO:297, and a light chain variable region comprising an amino acid sequence of SEQ ID NO:298 or amino acid sequences 90% identical thereto. 
     
     
         33 . The use of any one of  claims 30-32 , wherein the symptoms of fibrosis are one or more of pulmonary hypertension, right-sided heart failure, respiratory failure, hypoxia, cough, formation of blood clots, pneumonia, and/or lung cancer in the subject. 
     
     
         34 . The use of any one of  claims 30-33 , wherein the subject has one or more risk factors for fibrosis selected from the group consisting of cigarette smoking, environmental factors and genetic predisposition for lung fibrosis. 
     
     
         35 . The use of any one of  claims 30-34 , further comprising the steps of:
 (i) determining the levels of circulating latent TGFβ in the subject prior to administering the TGFβ antibody or antigen-binding fragment thereof; and   (ii) determining the levels of circulating latent TGFβ in the subject after administering the TGFβ antibody or antigen-binding fragment thereof,   wherein an increase in circulating latent TGFβ after antibody or antigen-binding fragment thereof administration, as compared to circulating latent TGFβ before administration, indicates therapeutic efficacy.   
     
     
         36 . The use of any one of  claims 31-35 , wherein reduction in the amount of collagen present in the fibrotic tissue, reduction in the amount of new collagen synthesis, and/or reduction in the amount of phosphorylated Smad2 in the fibrotic tissue is determined 24 hours, 48 hours, 72 hours, or 96 hours after administration of the TGFβ inhibitor. 
     
     
         37 . The use of any one of  claims 31-36 , further comprising a step of selecting a subject who would benefit from a reduction in a level of collagen, a level of new collagen synthesis, and/or a level of phosphorylated Smad2 in a fibrotic tissue. 
     
     
         38 . The use of any one of  claims 30-37 , wherein the loading dose/maintenance dose regimen comprises a loading dosage of between about 30 mg/kg and about 90 mg/kg and a maintenance dosage of between about 10 mg/kg and about 30 mg/kg. 
     
     
         39 . The use of  claim 38 , wherein the loading dosage is about 30 mg/kg and the maintenance dosage is about 10 mg/kg. 
     
     
         40 . A TGFβ1 inhibitor for use in the treatment of a cancer associated with elevated levels of reactive oxygen species (ROS) in a subject, wherein the treatment comprises administering the TGFβ1 inhibitor in an amount sufficient to the subject in need thereof, wherein the TGFβ1 inhibitor is an antibody or antigen-binding fragment thereof, optionally a TGFβ1-selective antibody or antigen-binding fragment thereof. 
     
     
         41 . The TGFβ1 inhibitor for use of  claim 40 , wherein the elevated levels of ROS are associated with a genotoxic therapy. 
     
     
         42 . The TGFβ1 inhibitor for use of  claim 40 or 41 , wherein the subject is further treated with a checkpoint inhibitor. 
     
     
         43 . A TGFβ1 inhibitor for use in the treatment of immune suppression in a subject, wherein the treatment comprises administration of a TGFβ1 inhibitor to a patient treated with a genotoxic therapy, wherein optionally the subject has cancer, and wherein further optionally the genotoxic therapy comprises a chemotherapy and/or radiation therapy. 
     
     
         44 . The TGFβ1 inhibitor for use according to  claim 43 , wherein the subject is further treated with a checkpoint inhibitor, wherein optionally the checkpoint inhibitor is an anti-PD-1 or anti-PD-L1 antibody.